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Abstract The study investigated the effect of soilless media (burlap), on the morphological traits and antioxidant activities of microgreens from Brassicaceae, Amaranthaceae, and Linaceae families. The results revealed significant variations were observed in the selected morphological, biochemical composition, and antioxidant capacity of the microgreens. The radish sango and microgreens showed superior morphological characteristics compared to other microgreens. The elemental composition analysis revealed consistent moisture, ash, fat, fiber, and protein content across all families. The results revealed significant variations in the biochemical composition and antioxidant capacity of the microgreens, depending on the growing medium and between microgreens. Notably, microgreens differed in photosynthetic pigment profiles, with flaxseed and cabbage showing the highest chlorophyll content of 26.59 to 27.18 µg/g, FW and carotenoid content in a range of 3.74 to 6.39 µg/g, FW was observed in microgreens. The radish sango and beetroot microgreens exhibited elevated anthocyanin levels of 27.94–28.25 µmol/100 g, FW. Biochemical analysis indicated varying levels of ascorbic acid (177.58 to 256.46 mg/100 g, FW), total glucosinolate content (4.09 to 47.38 µmol/g, FW), phenolic content (131.44 to 298.56 mg GAE/100 g, FW), and flavonoid content (10.94 to 18.14 mg QUE/100 g, FW) were observed in selected microgreens families. Radish sango microgreens demonstrated the highest DPPH (76.82%, FW) and ABTS (88.49%, FW) radical scavenging activities, indicating superior antioxidant potential. The study showed that Brassicaceae microgreens are particularly rich in bioactive and antioxidant properties. Additionally, studies could assess the economic feasibility and scalability of soilless cultivation methods for microgreens to support their inclusion in sustainable agricultural practices and health-promoting diets.

Corn silk (Zea mays L.), an abundant agricultural waste, contains various bioactive compounds that exhibit promising health benefits. The current study focuses on development and optimization of corn silk-based instant mix using response surface methodology. The optimized product, with 14.66% corn silk, 10% sugar and 0.22% xanthan gum in a skim milk powder base, scored 0.925 desirability. The physico-chemical and sensory parameters of optimized mix closely aligned with expected values. The instant mix packaged in metallised polyester yielded superior preservation of quality indicators over 120 days compared to low-density polyethylene (LDPE) and high-density polyethylene (HDPE). The microbial load in corn silk instant mix was observed across packaging materials and highlighting hydroxyl methyl furfural (HMF) as the primary predictor of product stability, the study calculated a 94.95 days half-life at 10 °C. Corn silk's rich bioactive compound supports its integration into nutraceuticals and instant mixes, mitigating food waste while enhancing nutritional value.Novelty statement.In this study, corn silk powder was utilized for the development of the instant mix. This innovative approach transforms corn silk, typically discarded as agricultural waste, into a commercially sustainable product that delivers the nutrients of corn silk to a broader population. Despite fresh corn silk being a perishable commodity, it has very low storage shelf life. The developed instant mix effectively preserves its nutritional value for up to six months, offering a sustainable and nutritious option for consumers.

Green leafy vegetables, especially microgreens are gaining popularity due to their high nutritional profiles, rich phytochemical content, and intense flavors. This review explores the growing commercial market for microgreens, especially in upscale dining and premium grocery outlets, highlighting consumer perceptions and their effect on market dynamics. Apart from these, the effect of modern agricultural methods that maximize the growth of microgreens is also examined. The value is anticipated to increase significantly, according to market predictions, from $1.7 billion in 2022 to $2.61 billion by 2029. Positive consumer views on microgreens health benefits drive this growth, although challenges such as varying levels of consumer awareness and income disparities affect sales. The review underscores the need for targeted research and strategic initiatives to enhance consumer understanding and improve cultivation methods to support market expansion in upcoming years.

Baby corn, characterized by its high water activity and elevated respiration rate, poses a formidable obstacle to prolonged storage under standard ambient conditions and necessitates specialized treatments for transportation to distant locations. One of the primary postharvest challenges associated with baby corn is the occurrence of brown pigment formation because of enzymatic browning at the apex of its immature ovules, cut surfaces, and silk attached to the young ears. The present study was undertaken to investigate the effect of different blanching treatments on peroxidase inactivation, physicochemical properties, and functional properties of baby corn. The treatments applied were hot water blanching (HWB) at temperatures ranging from 70°C to 90 °C for 30–240 s, steam blanching (SB) for 30–240 s, and microwave blanching (MWB) at power levels of 360 W–900 W for 30–300 s. Results indicated that 90 % peroxidase enzyme inactivation occurred under different methods as 90 °C for 60 s for HWB, 100 °C for 60 s for SB, and 540 W for 30 s for MWB. These blanching methods have shown significant effects on the properties under investigation. MWB demonstrated the highest retention of ascorbic acid (94.15 %) and minimal color changes (ΔE = 5.72) in comparison to hot water and steam blanching. Similarly, the result for total flavonoid content for 540 W, 90 °C and 100 °C for 30, 60, and 60 s were found to be 3.01,1.99 and 2.10 mg QE/100g, phenols 48.98, 47.99 and 48.03 mg GAE/100g and DPPH (%) 42.55, 34.20 and 37.08 % respectively. The findings suggest that microwave blanching of baby corn at 540 W for 30 s holds promise to inactivate the peroxidase enzyme with better retention of physicochemical and functional properties.

Bee pollen being superfood was incorporated in skim milk and spray-dried to obtain bee pollen enriched milk powder (BPEMP), which was stored in ALP, BOPP/LDPE, and PET/Al foil/LDPE pouches (commonly used to pack food powders) at 5, 25 and 40°C for 1 year. The current study aims the determination and comparison of the physico-chemical, phytochemical, and microstructural properties of BPEMP with control during storage. Water activity and peroxide value increased whereas there was a significant (p

Abstract The present research study aimed to examine three different herb extract's effects on the discoloration rate of fresh-cut pear slices using an image analysis technique. Pear slices were sprayed and dip-coated with Ocimum basilicum, Origanum vulgare, and Camellia sinensis (0.1 g/ml) extract solution. During 15 days storage period with three days intervals, all sprayed/dip-coated pear slices were analyzed for the quality attribute (TA) and color parameters notably a*, b*, hue angle (H*), lightness (L*), and total color change (ΔE). Further, order kinetic models were used to observe the color changes and to predict the shelf-life. The results obtained showed that the applicability of image analysis helped to predict the discoloration rate, and it was better fitted to the first-order (FO) kinetic model (R2 ranging from 0.87 to 0.99). Based on the kinetic model, color features ΔE and L* was used to predict the shelf-life as they had high regression coefficient values. Thus, the findings obtained from the kinetic study demonstrated Camellia sinensis (assamica) extract spray-coated pear slices reported approximately 28.63- and 27.95-days shelf-stability without much discoloration compared with all other types of surface coating.

ABSTRACTThe objective of this study was to detect the changes in physicochemical and functional properties of raw and processed divergent honey varieties after adulterating with different sugar syrups. Specifically, when 25% to 55% cane syrup was added to raw Acacia honey, the HMF content increased from 46.25 to 101.6 mg/kg. In processed honey that contained 55% cane syrup, HMF content was even higher, reaching up to 402.47 mg/kg. The reported values for HMF (>80 mg/kg) content and DN (>8 DN) were above the described quality evaluation standard of honey. The results indicated that adulterating Acacia honey with up to 55% corn and corn syrup caused a reduction in diastase content to below 3.5 DN, and an increase in HMF content above 93.28 mg/kg. Similarly, adding 10% corn syrup to raw Ziziphus honey resulted in a decrease in both DPPH and ABTS+ values from 83.16% and 88.58%, respectively, to 72.83% and 76.97%. The findings in this study demonstrated that the addition of malt syrup (55%) to raw Trifolium honey resulted in an increase in the phytochemical content, with the TPC value reaching 805.15 mg GAE/100 g. However, processing Trifolium honey caused a significant decrease in TPC content to 505.15 mg GAE/100 g. This work highlights the need for more research into unexplored honey varieties and adulteration methods to improve quality evaluation standards and mitigate authenticity issues.

Collagens are the most abundant structural proteins in the extracellular matrix of animals and play crucial roles in maintaining the structural integrity and mechanical properties of tissues and organs while mediating important biological processes. Fibrillar collagens have a unique triple helix structure with a characteristic repeating sequence of (Gly-X-Y)n. Variations within the repetitive sequence can cause misfolding of the triple helix, resulting in heritable connective tissue disorders. The most common variations are single-point missense mutations that lead to the substitution of a glycine residue with a bulkier amino acid (Gly → X). In this review, we will first discuss the importance of collagen’s triple helix structure and how single Gly substitutions can impact its folding, structure, secretion, assembly into higher-order structures, and biological functions. We will review the role of “designer collagens,” i.e., synthetic collagen-mimetic peptides and recombinant bacterial collagen as model systems to include Gly → X substitutions observed in collagen disorders and investigate their impact on structure and function utilizing in vitro studies. Lastly, we will explore how computational modeling of collagen peptides, especially molecular and steered molecular dynamics, has been instrumental in probing the effects of Gly substitutions on structure, receptor binding, and mechanical stability across multiple length scales.

Molecular level control is required to capture the folding and supramolecular assembly of collagen in mimetic materials. Here, the authors report on the creation of a synthetic collagen which assembles into banded fibers, recaptures structural properties of natural collagen and which can act as a testbed for design and experimentation

Streptococcus pyogenes-derived recombinant bacterial collagen-like proteins (CLPs) are emerging as a potential biomaterial for biomedical research and applications. Bacterial CLPs form stable triple helices and lack specific interactions with human cell surface receptors, thus enabling the design of novel biomaterials with specific functional attributes. Bacterial collagens have been instrumental in understanding collagen structure and function in normal and pathological conditions. These proteins can be readily produced in E. coli, purified using affinity chromatography, and subsequently isolated after cleavage of the affinity tag. Trypsin is a widely used protease during this purification step since the triple helix structure is resistant to trypsin digestion. However, the introduction of Gly→X mutations or natural interruptions within CLPs can perturb the triple helix structure, making them susceptible to trypsin digestion. Consequently, removing the affinity tag and isolating collagen-like (CL) domains containing mutations is impossible without degradation of the product. We present an alternative method to isolate CL domains containing Gly→X mutations utilizing a TEV protease cleavage site. Protein expression and purification conditions were optimized for designed protein constructs to achieve high yield and purity. Enzymatic digestion assays demonstrated that CL domains from wild-type CLPs could be isolated by digestion with either trypsin or TEV protease. In contrast, CLPs containing Gly→Arg mutations are readily digested by trypsin while digestion with TEV protease cleaved the His6-tag, enabling the isolation of mutant CL domains. The developed method can be adapted to CLPs containing various new biological sequences to develop multifunctional biomaterials for tissue engineering applications.

The aim of the study was to characterize four unique varieties of honey (saffron, apple, cherry and Plectranthus rugosus) from Kashmir valley of India on the basis of physico-chemical parameters (moisture content, pH, hydroxymethylfurfural, water activity, proline content, invertase and diastase activity) and carbohydrate profile. Also the effect of temperature (0, 5, 10, 20 and 30 °C) on rheological properties of all unique honey varieties was studied. All the physicochemical parameters studied were significantly different (P

Iron-sulfur proteins are primordial catalysts and biological electron carriers that today drive major metabolic pathways across all forms of life. They can access a diversity of oxidation states and can mediate electron transfer over an extended range of reduction potentials spanning more than 1 V. Depending on the protein micro-environment and geometry of ligand, co-ordination the iron-sulfur clusters can occur in different forms [2Fe-2S], [3Fe-4S], HiPIP [4Fe-4S], and [4Fe-4S]. There are several spectroscopic methods available to characterize the composition and electronic configuration of the iron-sulfur clusters, such as optical methods and electron paramagnetic resonance. This paper presents the protocols used to characterize the metal center of Coiled-Coil Iron-Sulfur (CCIS), an artificial metalloprotein containing one [4Fe-4S] cluster. It is expected that these protocols will be of general utility for other iron-sulfur proteins.

Iron-sulfur proteins are ubiquitous among all living organisms and are indispensable for almost all metabolic pathways ranging from photosynthesis, respiration, nitrogen, and carbon dioxide cycles. The iron-sulfur clusters primarily serve as electron acceptors and donors and transfer electrons to active sites of various enzymes, thus driving the energy metabolism. Prokaryotes like E. coli have ISC and SUF pathways that help in the assembly and maturation of iron-sulfur proteins. These iron-sulfur proteins, especially with [4Fe-4S] clusters, are highly sensitive to molecular oxygen, and it would be advantageous if the de novo proteins and native proteins having iron-sulfur binding sites are expressed and isolated under anaerobic conditions. Bacterially assembled iron-sulfur proteins, when isolated and purified anaerobically, exhibit improved biochemical and biophysical stabilities in comparison to the counterparts expressed and purified aerobically and reconstituted under anaerobic conditions. This protocol outlines the expression and purification of the artificial protein, Coiled-Coil Iron-Sulfur (CCIS). It may be deployed to both natural and artificial [4Fe-4S] proteins when heterologously expressed in E. coli.

Objective: The present study assessed physico-chemical, techno-functional, thermal and textural characteristics of propolis samples (n = 30) collected from four Northern zones of India. Methods: The propolis samples were analyzed according to the Association of Official Analytical Chemists (AOAC) official protocols for their ash, moisture, and protein content. Soxtech, Fiber tech, Hunter LAB Colorimeter, TA.XT2i Texture Analyzer, Thermo gravimetric Analyzer, Discovery DSC 25 have been used to assess the crude fat, fibre content, colour, texture and thermal properties, respectively. Results and conclusions: Physico-chemical properties of propolis differed significantly (p

Objective: The study was carried out to investigate the rheological behavior and storage studies of sprouted onion pastes from four varieties (Punjab White, Punjab Naroya, PRO-6 and commercial). Methods: The storage study of paste samples from raw and sprouted onions packaged in polyethylene terephthalate (PET) bottles was carried out at room temperature (25 ± 1 °C) after thermally processing them at 80 °C for 15 min. All the paste samples were periodically analyzed for the change in quality characteristics viz, colour, pH, titratable acidity, TSS(∘B), TS (%), Water activity, Moisture (%), Total phenol content (mg/g), Total flavonoid content (mg/g), Total antioxidant capacity (mg/g), DPPH (%) and microbial count. The rheological properties of raw and sprouted onion paste samples were studied using Modular Compact Rheometer MCR-102 equipped with parallel plates geometry. Results and conclusions: Sprouted onion pastes were found to retain quality characteristics with lesser color changes, higher total phenolics, total flavonoids, antioxidant activity and microbial stability than raw pastes during storage. Storage period and its interactive effect with the variety and sprouting had significant (P

Objective: This study was carried out to demonstrate the effect of sprouting on the major flavonols and quercetin glycosides in onion powders developed from four onion cultivars (Punjab White, Punjab Naroya, PRO-6 and Commercial) of Indian Origin. Methods: The major flavonols summing up total flavonoids and quercetin glucosides comprising up to the total quercetin glucoside concentration were determined by High performance liquid chromatography and Fourier transform infrared (FTIR) spectroscopy. Results and conclusions: The major flavonols were quercetin, myricetin and kaempferol, which were found to be significantly higher (p

Corn silk has long been thought of as a waste product; however, due to its numerous therapeutic attributes, it has remarkably gained popularity in Asian and African countries. Therefore, this study aimed to assess the bioactivity of dried corn silk powder (Zea mays, G5417) in terms of its physicochemical and bio-functional characteristics. The protein (15.29 ± 1.23) and ash (5.29 ± 0.29) contents in the corn silk powder were found to be high. The high phenolic content (94.10 ± 0.26 mg GAE/g) and flavonoid content (163.93 ± 0.83 mg QE/100 g) are responsible for its high antioxidant activity. The corn silk powder showed 45.40 ± 0.92% FRSA, 75.25 ± 0.59 TEAC mg/gdw of ABTS, and 86.77 ± 0.88% of FRAP. FT-IR spectroscopy revealed stretching, bending, and vibrations of abundantly present polysaccharides and protein functional groups. Moreover, the DSC thermograph revealed the exothermic reactions at on-set temperature (Tonset) = 21.9 °C and end temperature (Tendset) = 102.80 °C, and exothermic reactions at on-set temperature (Tonset) = 252.02 °C, end temperature (Tendset) = 296.80 °C, and denaturation peak temperature (Tpeak) = 277.48 °C, whereas XRD (2θ = 21.5°) confirmed the amorphous nature of the corn silk powder. Therefore, due to the potential bioactivity and thermal stability, dry corn silk powder can be scaled up at an industrial level.

Novel food packaging technologies arose as a result of consumer’s desire for convenient, ready to eat, tasty and mild processed food products with extended shelf life and maintained quality. Recent trend of lifestyle changes with less time for consumers to prepare foods posed a great challenge toward food packaging sector for the evolution of novel and innovative food packaging techniques. The novel food packaging techniques, viz. active packaging, intelligent packaging and bio active packaging which involve intentional interaction with the food or its surroundings and influence on consumer’s health have been the major innovations in the field of packaging technology. These novel techniques act by prolonging the shelf life, enhancing or maintaining the quality, providing indication and to regulate freshness of food product. The advancement in novel food packaging technologies involves retardation in oxidation, hindered respiratory process, prevention of microbial attack, prevention of moisture infusion, use of CO2 scavengers/emitters, ethylene scavengers, aroma emitters, time-temperature sensors, ripeness indicators, biosensors and sustained release of antioxidants during storage. The novel food packaging technologies besides the basic function of containment increase the margin of food quality and safety. The novel food packaging techniques thus help in fulfilling the demands throughout the food supply chain by gearing up toward persons own lifestyle. The main objectives of this review article are to provide basic knowledge of different new and innovative food packaging techniques about their way of preservative action, effectiveness and suitability in various types of foods. Keywords: Innovative, Shelf life, Packaging, Bio active

The rheological properties of high altitude India honeys were analyzed in honeydew (pine) and nectar honey (multifloral and acacia) varieties at a wide range of temperatures (0, 5, 10, 20, and 30 °C). All the honey samples were significantly dominated by loss modulus (G″) which displayed their viscous nature. Irrespective of geographical origin and temperature, all the honey varieties showed a Newtonian behavior. The viscosity of all honey varieties showed a strong dependence on temperature, and Arrhenius model was examined to describe this. Keywords: Honey, Loss modulus, Storage modulus, Viscosity, Arrhenius model

Honey is a natural product well known for its nutritive value and being usually used as natural sweetener. India is one of the major honey exporting countries in the world. The present research refers to an investigation of the physico chemical and rheological behavior of four freshly harvested unifloral honey samples collected from different plant sources viz Sunflower (H1), Eucalyptus (H2), Mustard (H3) and Prosopis (H4). Rheological studies were carried out by varying honey samples temperature from 10 to 50 oC. Three different models (Arrhenius, Ostwald-de Waele Power law and Newtonian-I) were investigated. A continuous fall in apparent viscosity (p0.05) in apparent viscosity of honey was observed with the variation in shear rate, indicating their Newtonian behavior. The moisture content present in different honey samples (H1, H2, H3 and H4 was 13.6 %, 17.2 %, 15.5 % and 16.1 % respectively) also had a significant effect (p 99%) in explaining the rheological behavior of honey following the variations in temperature when compared with Arrhenius and Newtonian-I respectively.

The study was intended to characterize three honeys (acacia, pine honeydew and multifloral) from high altitude Kashmir valley of India according to their macro minerals (K, Ca, Na and P), antioxidant properties and sugar parameters. The result for total phenolic content (22.68–59.84 mg GAE/100 g) and total flavonoid content (6.10–8.12 mg QE/100 g), revealed that honeys from Kashmir valley have high antioxidant activity. Principal component analysis (PCA), explained more than 81% of the variance. Four sugars were identified and quantified by HPLC, which include monosaccharides and disaccharides. Chemometric methods such as principal component analysis and linear discriminate techniques were applied on the data in order to differentiate the honeys. PCA explained more than 81% of the variance with the first two PC variables with minerals and antioxidant properties having highest discriminating power while LDA successfully classified all the unifloral honey samples. Keywords: Honey, Antioxidant, Minerals, PCA, LDA

The present study was done to optimize the power ultrasound processing for maximizing diastase activity of and minimizing hydroxymethylfurfural (HMF) content in honey using response surface methodology. Experimental design with treatment time (1-15 min), amplitude (20-100 %) and volume (40-80 mL) as independent variables under controlled temperature conditions was studied and it was concluded that treatment time of 8 min, amplitude of 60 % and volume of 60 mL give optimal diastase activity and HMF content, i.e. 32.07 Schade units and 30.14 mg/kg, respectively. Further thermal profile analyses were done with initial heating temperatures of 65, 75, 85 and 95 ºC until temperature of honey reached up to 65 ºC followed by holding time of 25 min at 65 ºC, and the results were compared with thermal profile of honey treated with optimized power ultrasound. The quality characteristics like moisture, pH, diastase activity, HMF content, colour parameters and total colour difference were least affected by optimized power ultrasound treatment. Microbiological analysis also showed lower counts of aerobic mesophilic bacteria and in ultrasonically treated honey than in thermally processed honey samples complete destruction of coliforms, yeasts and moulds. Thus, it was concluded that power ultrasound under suggested operating conditions is an alternative nonthermal processing technique for honey.

The range of barrel-shaped proteins found in the outer membrane of certain bacteria evolved through multiple pathways.

In the present study, the spray-dried honey powder enriched with aonla (Emblica officinalis Gaertn) and basil (Ocimum sanctum) extract was developed using drying aids—gum arabic (GA), maltodextrin (MD), and whey protein concentrate (WPC), and then characterized based on particle size distribution, colour, glass transition temperature (Tg), X-ray diffraction, and antioxidant and rheological properties. Results showed the highest Tg (86.13 °C) for WPC based honey powder, which, in turn, resulted in least stickiness as compared to GA and MD based honey powders with Tg 74.53 °C and 68.26 °C, respectively. The dried honey powder with all three carrier agents exhibited a metastable amorphous state as proved by the broader peaks of X-ray diffractograms due to the short drying time, whereas, a peak at 1637 cm−1, attributed to the carbonyl (C=O) stretching, established the ascorbic acid in the studied powder on account of aonla extract. The mean particle diameter significantly (p < 0.05) increased, following the order WPC (60.45 μm) > GA (41.24 μm) > MD (20.06 μm) as carrier agents, which were related to the higher feed viscosity. The colour parameter L* (30.74–45.78) and b* (5.82–11.64) values of the nutritionally rich honey powder were higher due to presence of polyphenols in aonla and basil extracts, which resulted in the formation of dark brown complexes. The antioxidant activity of WPC based fortified honey powder was highest (82.73%), followed by GA (78.15%) and MD (74.85%) based honey powders. A significant (p < 0.05) increase was found in powder recovery, solubility, and dispersibility using the drying aids in the following order: WPC < GA < MD. Furthermore, the storage modulus (G′) was higher than loss modulus (G″) in all honey powders, wherein the WPC containing powder demonstrated maximum value of G′, followed by GA and MD. Finally, the three honey powders were microbiologically stable.

The morphology and severity of human congenital cataract varies even among individuals with the same mutation, suggesting that genetic background modifies phenotypic penetrance. The spontaneous mouse mutant, vacuolated lens (vl), arose on the C3H/HeSnJ background. The mutation disrupts secondary lens fiber development by E16.5, leading to full penetrance of congenital cataract. The vl locus was mapped to a frameshift deletion in the orphan G protein-coupled receptor, Gpr161, which is expressed in differentiating lens fiber cells. When Gpr161vl/vl C3H mice are crossed to MOLF/EiJ mice an unexpected rescue of cataract is observed, suggesting that MOLF modifiers affect cataract penetrance. Subsequent QTL analysis mapped three modifiers (Modvl3-5: Modifier of vl) and in this study we characterized Modvl4 (Chr15; LOD = 4.4). A Modvl4MOLF congenic was generated and is sufficient to rescue congenital cataract and the lens fiber defect at E16.5. Additional phenotypic analysis on three subcongenic lines narrowed down the interval from 55 to 15Mb. In total only 18 protein-coding genes and 2 micro-RNAs are in this region. Fifteen of the 20 genes show detectable expression in the E16.5 eye. Subsequent expression studies in Gpr161vl/vl and subcongenic E16.5 eyes, bioinformatics analysis of C3H/MOLF polymorphisms, and the biological relevancy of the genes in the interval identified three genes (Cdh6, Ank and Trio) that likely contribute to the rescue of the lens phenotype. These studies demonstrate that modification of the Gpr161vl/vl cataract phenotype is likely due to genetic variants in at least one of three closely linked candidate genes on proximal Chr15.

The purpose of the present work was to study the effect of inlet temperature (160–180°C), feed rate (0.08–0.13 ml/s), concentration of gum Arabic (35–45%), aonla extract (6–8%), and basil extract (6–8%) on the product properties (bulk density, hygroscopicity, total phenolic content (TPC), antioxidant activity (AOA), and vitamin C content) of spray-dried nutritionally rich honey powder using response surface methodology. Higher inlet air temperatures led to lower bulk density and hygroscopicity, whereas addition of aonla and basil extracts led to higher TPC, AOA, and vitamin C content which were encapsulated by gum Arabic. Statistical analysis showed that independent variables significantly affected all the responses (p

Multiplexed cloning of long DNA sequences is a valuable technique in many biotechnology applications, such as long-read genome sequencing and the creation of open reading frame (ORF) libraries. Long-adapter single-stranded oligonucleotide (LASSO) probes have shown promise as a tool to clone long DNA fragments. LASSO probes are molecular inversion probes (MIP) engineered with an adapter region of user-defined length, flanked between template-specific probe sequences. Herein, we demonstrate that the adapter length is a key feature of LASSO that influences the efficiency of gene capture and cloning. Furthermore, we applied a model based on Monte Carlo molecular simulation in order to study the relationship between the long-adapter length of LASSO and capture enrichment. Our results suggest that the adapter length is a factor that contributes to the free energy of target−probe interaction, thereby determining the efficiency of capture. The results indicate that LASSOs with extremely long adapters cannot capture the targets well. They also suggest that targets of different lengths may prefer adapters of different lengths.

Charge-pair interactions between acidic and basic residues on the surface of collagen can promote stability as well as control specificity of molecular recognition. Heterotrimeric collagen peptides have been engineered de novo using either rational or computational methods, which in both cases optimize networks of favorable charge-pair interactions in the target structure. Less understood is the role of electrostatic repulsion between groups of like charge in destabilizing structure or directing molecular recognition. To study this, we apply a “charge crowding” approach, where repulsive interactions between multiple aspartate side chains are found to destabilize the homotrimer states in triple helical peptide system and can be utilized to promote the formation of heterotrimers. Neutralizing surface charge by increasing salt concentration or decreasing pH can enhance homotrimer stability, confirming the role of charge crowding on the destabilization of homotrimers via electrostatic repulsion. Charge crowding may be used in conjunction with other approaches to create specific collagen heterotrimers.

Recently, catalytic peptides were introduced that mimicked protease activities and showed promising selectivity of products even in organic solvents where protease cannot perform well. However, their catalytic efficiency was extremely low compared to natural enzyme counterparts presumably due to the lack of stable tertiary fold. We hypothesized that assembling these peptides along with simple hydrophobic pockets, mimicking enzyme active sites, could enhance the catalytic activity. Here we fused the sequence of catalytic peptide CP4, capable of protease and esterase-like activities, into a short amyloidogenic peptide fragment of Aβ. When the fused CP4-Aβ construct assembled into antiparallel β-sheets and amyloid fibrils, a 4.0-fold increase in the hydrolysis rate of p-nitrophenyl acetate (p-NPA) compared to neat CP4 peptide was observed. The enhanced catalytic activity of CP4-Aβ assembly could be explained both by pre-organization of a catalytically competent Ser-His-acid triad and hydrophobic stabilization of a bound substrate between the triad and p-NPA, indicating that a design strategy for self-assembled peptides is important to accomplish the desired functionality.

With increasing consumers demand and tightening of food and environmental regulations, traditional food-processing techniques have lost their optimum performance which gave rise to new and powerful technologies. Ultrasonic is a one of the fast, versatile, emerging, and promising non-destructive green technology used in the food industry from last few years. The ultrasound is being carried out in various areas of food technology namely crystallization, freezing, bleaching, degassing, extraction, drying, filtration, emulsification, sterilization, cutting, etc. Ultrasound is being applied as an effective preservation tool in many food-processing fields viz. vegetables and fruits, cereal products, honey, gels, proteins, enzymes, microbial inactivation, cereal technology, water treatment, diary technology, etc. This review summarizes the latest knowledge on impact and application of ultrasound in food technology.

Use of fusion protein tags to investigate lysosomal proteins can be complicated by the acidic, protease-rich environment of the lysosome. Potential artifacts include degradation or release of the tag and acid quenching of fluorescence. Tagging can also affect protein folding, glycosylation and/or trafficking. To specifically investigate the use of fluorescent tags to reveal lysosomal localization, we tested mCherry derivatives as C-terminal tags for Niemann-Pick disease type C protein 2 (NPC2), a luminal lysosomal protein. Full-length mCherry was released from the NPC2 chimera while deletion of the 11 N-terminal residues of mCherry generated a cleavage-resistant (cr) fluorescent variant. Insertion of proline linkers between NPC2 and crmCherry had little effect while Gly-Ser linkers promoted cleavage. The NPC2-crmCherry fusion was targeted to the lysosome and restored function in NPC2-deficient cells. Fusion of crmCherry to known and candidate lysosomal proteins revealed that the linkers had different effects on lysosomal localization. Direct fusion of crmCherry impaired mannose 6-phosphorylation and lysosomal targeting of the lysosomal protease tripeptidyl peptidase I (TPP1), while insertion of linkers corrected the defects. Molecular modeling suggested structural bases for the effects of different linkers on NPC2 and TPP1 fusion proteins. While mCherry fusion proteins can be useful tools for studying the lysosome and related organelles, our findings underscore the potential artifacts associated with such applications.

Visualizations that are low-cost in memory are desirable. We present a method for stitching three-dimensional scattered data from multiple worksheets into a dynamic “animation-like” surface chart in Excel. This method is useful when (1) the user hard-codes the data points to conserve memory; employing such strategy scales better than soft-coding data values, (2) the data values are hard-coded by an unknown source, or (3) the function is complex and requires a user-defined function to output values into cells. In particular, we demonstrate an application in biology where rigid motion (rotation and translation are the only transformations applied to an object in 3-D space) is used to model the free energy gain/loss by surveying various placements and orientations of membrane proteins with respect to their environment. Our strategy involves a simple concept of scrolling through an order of worksheets, and can be extended to even more dimensions (i.e. scrolling through workbooks if necessary)

Models of early protein evolution posit the existence of short peptides that bound metals and ions and served as transporters, membranes or catalysts. The Cys-X-X-Cys-X-X-Cys heptapeptide located within bacterial ferredoxins, enclosing an Fe₄S₄ metal center, is an attractive candidate for such an early peptide. Ferredoxins are ancient proteins and the simple α+β fold is found alone or as a domain in larger proteins throughout all three kingdoms of life. Previous analyses of the heptapeptide conformation in experimentally determined ferredoxin structures revealed a pervasive right-handed topology, despite the fact that the Fe₄S₄ cluster is achiral. Conformational enumeration of a model CGGCGGC heptapeptide bound to a cubane iron-sulfur cluster indicates both left-handed and right-handed folds could exist and have comparable stabilities. However, only the natural ferredoxin topology provides a significant network of backbone-to-cluster hydrogen bonds that would stabilize the metal-peptide complex. The optimal peptide configuration (alternating α(L),α(R)) is that of an α-sheet, providing an additional mechanism where oligomerization could stabilize the peptide and facilitate iron-sulfur cluster binding.